摘要
目的探究芥子碱硫氰酸盐(sinapine thiocyanate)对肝癌细胞SMMC-7721增殖、迁移和侵袭的影响及机制。方法采用CCK-8、划痕和Transwell实验,检测不同浓度(0、25、50和100μmol·L^-1)的芥子碱硫氰酸盐对SMMC-7721细胞增殖、迁移和侵袭能力的影响;利用生物信息学和分子对接技术,挖掘蛋白靶点并验证芥子碱硫氰酸盐与蛋白靶点PTGS1和PTGS2的结合能力;Western blot检测芥子碱硫氰酸盐对SMMC-7721细胞中PTGS1、PTGS2、Bcl-2、Bax、MMP-2和MMP-9蛋白表达水平的影响。结果芥子碱硫氰酸盐明显抑制SMMC-7721细胞的增殖、迁移和侵袭的能力(P<0.05);芥子碱硫氰酸盐有27个蛋白靶点,蛋白靶点富集在氮代谢等4个通路上;其中,PTGS1和PTGS2富集在多个信号通路中,且其活性口袋能与芥子碱硫氰酸盐稳定结合;芥子碱硫氰酸盐明显减少肝癌细胞SMMC-7721中PTGS1、PTGS2、Bcl-2、MMP-2和MMP-9的蛋白表达(P<0.05),增加Bax的蛋白表达(P<0.05)。结论芥子碱硫氰酸盐靶向PTGS1和PTGS2,从而抑制肝癌细胞SMMC-7721增殖、迁移和侵袭。
Aim To investigate the effects of sinapine thiocyanate on the proliferation,migration and invasionof hepatocellular carcinoma SMMC-7721 cells and its mechanism.Methods CCK-8,wound healing and Transwell assays were performed to detect the effects of different concentrations(0,25,50 and 100μmol·L^-1)of sinapine thiocyanate on the proliferation,migration and invasion of SMMC-7721 cells.Bioinformatics technology and molecular docking technology were used to investigate protein targets of sinapine thiocyanate,and verify the binding between sinapine thiocyanate and its targets such as PTGS1 and PTGS2.Western blot was used to detect the effects of sinapine thiocyanate on the expression of PTGS1,PTGS2,Bcl-2,Bax,MMP-2 and MMP-9 in SMMC-7721 cells.Results Sinapine thiocyanate inhibited the proliferation,migration and invasion of SMMC-7721 cells(all P<0.05);Total 27 proteins were identified as the targets of sinapine thiocyanate,while they were enriched in four pathways including nitrogen metabolism pathway.Among them,the protein targets of sinapine thiocyanate,PTGS1 and PTGS2,were enriched in several signaling pathways,and their activated pocket could stably bound with sinapine thiocyanate.Western blot results showed that sinapine thiocyanate decreased the expression of PTGS1,PTGS2,Bcl-2,MMP-2 and MMP-9 in SMMC-7721 cells(P<0.05),as well as increased the expression levels of Bax(P<0.05).Conclusions Sinapine thiocyanate inhibits the proliferation,migration and invasion of human hepatocellular carcinoma SMMC-7721 cells via targeting PTGS1 and PTGS2.
作者
陈腾祥
王婧雅
曾智锐
雷珊
薛燕
孙远梅
杨宇石
兰金芝
CHEN Teng-xiang;WANG Jing-ya;ZENG Zhi-rui;LEI Shan;XUE Yan;SUN Yuan-mei;YANG Yu-shi;LAN Jin-zhi(Dept of Physiology,School of Basic Medical Science,Guizhou Provincial Key Lab of Pathogenesis&Drug Research on Common Chronic Diseases,Guizhou Medical University,Guiyang 550004,China;Dept of Pathology,Affiliated Hospital,Guizhou Medical University,Guiyang 550004,China;Dept of Pathology,Guizhou Medical University,Guiyang 550004,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2020年第12期1731-1737,共7页
Chinese Pharmacological Bulletin
基金
贵州省第五批人才基地建设项目[黔人领发(2016)22号]
黔科合平台人才[2018]5779-76
贵州医科大学研究生校级基金[贵医YJSCXJH(2019)002]。
